Development of unique instrumentation often using novel approaches is, in many instances, a necessary attribute to the success of biomedical research. Areas of emphasis are summarised below. Digital Video EEG Imaging System with Seizure Detection for Multiple Small Animals: The system, being developed in collaboration with Dr. Michael Rogawski, NINDS, will detect and record seizures of small animals by monitoring their EEG together with documenting their accompanying behavior via video recording. The instrumentation consists of animal cages, swivel connectors connected to the head of each animal, EEG analog amplifiers, computer control with movie maker and graphics hardware, and software detection of spikes and seizure patterns. It is presently being assembled and will visually display and record EEG data along with video of multiple small animals. Electronic Instrumentation for flash photolysis of Microsomal P450: In collaboration with Dr. Fred Friedman, NCI, a laser flash photolysis apparatus was developed to measure the kinetics of carbon monoxide (CO) binding to cytochromes P450 in liver microsomes from rats treated with various drugs and carcinogens. Since numerous forms of P450 contribute to the overall reaction, a difference kinetic method was used to distinguish the kinetic behavior of individual P450s. This method entails analysis of the difference between the kinetic profiles in the absence and presence of a specific P450 effector, and successfully yielded kinetic parameters for individual P450s involved in drug and carcinogen metabolism. Specifically, various polycyclic hydrocarbons differentially accelerated CO binding to the P450 1A1 which metabolizes these carcinogens in a size and shape dependent manner. The present acquisition and front panel control software is an application specific program which has no flexibility for investigator use. The proposed change in the acquisition, control, analysis and processing will be accomplished using the programming language Visual Basic together with the application program Component Works. In addition, analysis may include a semi automatic technique using mathematical processing software. Near-field scanning microscopy instrumentation: In a collaborative effort with Dr. Jeeseong Hwang, NIST, DBEPS is supporting Dr. James Dvorak and other NIAID investigators to implement a near-field scanning optical microscope in conjunction with atomic force microscopy. Excitation and emission optical pathways through an epi-configuration are being evaluated as an alternative approach to the conventional transmission methodology. Optical instrumentation for monitoring patients undergoing functional MRI evaluation: In this collaborative project with Dr. Andrew Goldfine, NINDS, patients with Tourette's syndrome are examined using MRI imaging to analyze activity within the brain. To allow correlation of these images with a visual record of the facial ticks and features, optical instrumentation was designed. The high magnetic field associated with the scanner required that illumination and video recording be performed remotely. Optical fibers were used for illumination. A custom designed non-magnetic concave mirror provided a full view of the patient?s face and video cameras with wide angle and telephoto lenses were employed for capturing the images tape. Modifications to Optical Motion Capture System for Clinical Gait Analysis: The system consists of video, imaging and electronic instrumentation for clinical gait analysis in the Rehabilitation Medicine Department of the Clinical Center. Included are a motion capture system, which synchronizes video data with force plates that measure ground reaction forces for kinetic analysis, EMG systems for muscle activity, and flexible modeling software to collect motion-tracking data. At present, the electronic ground system is being reconfigured in collaboration with Dr. Steven Stanhope, CC, in order to lower system noise. Photoprotection of tracheotomy intubation tubes: Nosocomial infection developed through tracheotomy intubation devices is an unwanted complication of patient management. Photo-irradiation of cells in proximity to singlet oxygen generating agents is widely known to cause cell death. Varied light delivery techniques and distribution of singlet oxygen generating dyes are being studied in collaboration with Dr. Theodor Kolobow, NHLBI, for efficacy in producing cell death. Enhancements to Small Animal Biophotonic Imaging System: The system accomplishes whole body optical imaging of small animals for assaying molecular and cellular processes as they occur in vivo. It non-invasively images reporter genes, such as luciferase and green fluorescent protein (GFP). For bioluminescent imaging, light is emitted from the animal after systemic administration of the substrate luciferin. Using GFP expressing cells, fluorescent noninvasive imaging employs an external light source of the appropriate wavelength to excite the fluorescent marker within the tissue of interest. Improved techniques in capture, processing and semi-automatic analysis of the images are being developed in collaboration with Dr. Steven Libutti, NCI. Small Animal CCD-Based Digital X-Ray Imaging System: The imaging system, recommended by DBEPS, will provide detailed anatomical, structural, and physiological function through high resolution imaging of small animals. Improved techniques in processing and analysis of the image will be developed in collaboration with Dr. Hynda Kleinman, NIDCR. These techniques will be applicable to a second fluorescent small animal imaging system that is under consideration.